Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy

Non-invasive genetic approaches, particularly using fecal samples, are commonly used to study endangered and elusive species, as they are easy to collect with minimal permission and cause little disturbance to the subject population. However, such studies face limitations due to poor DNA yield, whic...

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Main Authors: Jee Hyun Kim, Jangmi Lee, Dong Youn Kim, Yoon-Do Yang, Sujoo Cho, Han-Chan Park, Sung Yong Han, Mi-Sook Min, Hang Lee, Je-Yoel Cho, Puneet Pandey
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/15/4/574
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author Jee Hyun Kim
Jangmi Lee
Dong Youn Kim
Yoon-Do Yang
Sujoo Cho
Han-Chan Park
Sung Yong Han
Mi-Sook Min
Hang Lee
Je-Yoel Cho
Puneet Pandey
author_facet Jee Hyun Kim
Jangmi Lee
Dong Youn Kim
Yoon-Do Yang
Sujoo Cho
Han-Chan Park
Sung Yong Han
Mi-Sook Min
Hang Lee
Je-Yoel Cho
Puneet Pandey
author_sort Jee Hyun Kim
collection DOAJ
description Non-invasive genetic approaches, particularly using fecal samples, are commonly used to study endangered and elusive species, as they are easy to collect with minimal permission and cause little disturbance to the subject population. However, such studies face limitations due to poor DNA yield, which affects the overall utilization of collected samples and increases data errors. Here, we evaluated the impact of sample age and collection season on the performance of DNA extracted from feces (spraints) of the Eurasian otter (<i>Lutra lutra</i>), a semi-aquatic apex predator in South Korean freshwater ecosystems. We found that PCR amplification success rates decreased more rapidly in summer (79.3–58.2%) compared to winter (99.2–84.8%) with extended environmental exposure. Genotyping error rates were higher in samples collected during summer, with the rate of error increase over time being significantly greater in summer than in winter. The hot and humid South Korean summer fosters microbial growth and fecal degradation, which negatively impacts DNA yield, reducing PCR amplification success and increasing genotyping errors. We recommend collecting otter feces during winter for better DNA quality. If sampling in summer is unavoidable, it is crucial to collect fresh samples, which can be facilitated by conducting frequent surveys of latrine sites.
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spelling doaj-art-9b25a786537d4d1fbf7fac5c3c0eb4282025-08-20T03:11:03ZengMDPI AGAnimals2076-26152025-02-0115457410.3390/ani15040574Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data AccuracyJee Hyun Kim0Jangmi Lee1Dong Youn Kim2Yoon-Do Yang3Sujoo Cho4Han-Chan Park5Sung Yong Han6Mi-Sook Min7Hang Lee8Je-Yoel Cho9Puneet Pandey10Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Biological Sciences, Konkuk University, Seoul 05029, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaConservation Genome Resource Bank for Korean Wildlife, Seoul 08826, Republic of KoreaAssociation of Korean Otter Conservation, Hwacheon 24135, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Biochemistry, College of Veterinary Medicine, Research Institute for Veterinary Science, BK21 FOUR Future Veterinary Medicine Leading Education and Research Center, Seoul National University, Seoul 08826, Republic of KoreaResearch Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of KoreaNon-invasive genetic approaches, particularly using fecal samples, are commonly used to study endangered and elusive species, as they are easy to collect with minimal permission and cause little disturbance to the subject population. However, such studies face limitations due to poor DNA yield, which affects the overall utilization of collected samples and increases data errors. Here, we evaluated the impact of sample age and collection season on the performance of DNA extracted from feces (spraints) of the Eurasian otter (<i>Lutra lutra</i>), a semi-aquatic apex predator in South Korean freshwater ecosystems. We found that PCR amplification success rates decreased more rapidly in summer (79.3–58.2%) compared to winter (99.2–84.8%) with extended environmental exposure. Genotyping error rates were higher in samples collected during summer, with the rate of error increase over time being significantly greater in summer than in winter. The hot and humid South Korean summer fosters microbial growth and fecal degradation, which negatively impacts DNA yield, reducing PCR amplification success and increasing genotyping errors. We recommend collecting otter feces during winter for better DNA quality. If sampling in summer is unavoidable, it is crucial to collect fresh samples, which can be facilitated by conducting frequent surveys of latrine sites.https://www.mdpi.com/2076-2615/15/4/574non-invasive samplingEurasian otterDNA degradationPCR amplification successgenotyping errors
spellingShingle Jee Hyun Kim
Jangmi Lee
Dong Youn Kim
Yoon-Do Yang
Sujoo Cho
Han-Chan Park
Sung Yong Han
Mi-Sook Min
Hang Lee
Je-Yoel Cho
Puneet Pandey
Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
Animals
non-invasive sampling
Eurasian otter
DNA degradation
PCR amplification success
genotyping errors
title Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
title_full Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
title_fullStr Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
title_full_unstemmed Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
title_short Strategic Sampling of Eurasian Otter Spraints for Genetic Research in South Korea: Enhancing PCR Success and Data Accuracy
title_sort strategic sampling of eurasian otter spraints for genetic research in south korea enhancing pcr success and data accuracy
topic non-invasive sampling
Eurasian otter
DNA degradation
PCR amplification success
genotyping errors
url https://www.mdpi.com/2076-2615/15/4/574
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